Objective The purpose of this study was to evaluate the use of tracheo-oesophageal vioce prostheses after total laryngectomy.Methods Between 1995 and 1999,18 male patients ,Aged from 42 to 68,average 54.6,used Blom-singer voice prostheses after total laryngectomy.10 cases were glottic style,8 cases were subglottic style.12 patients received stage I,6 patients received stage Ⅱ voice reconstruction surgery. Results All cases were followed-up from 3 to 5 years. The success rate was 77.8%(14/18). The time with prostheses ranged from 8 to 19 months(average 12months). All failures were stage Ⅱ surgey .Conclusion Stage I voice reconstruction surgery can be performed in all total laryngectomy patients. But the indications of stage Ⅱ voice reconstruction surgery should be controlled strictly. More attention should be paid to the training of voice rehabilitation after total laryngectomy. The tracheo-oesophageal voice prostheses is an effective method to improve the life quality of the patients.

The triple negative breast cancer is one of importance in clinical subtypes of breast cancer, which is easy to recur and metastasis, and its prognosis is very poor.Radiotherapy, as an effective method for breast cancer,can reduce the risk of local recurrence.This article elaborates its characteristics,the progress of ra-diotherapy in the breast conserving surgery and modified radical mastectomy in triple negative breast cancer,when is the appropriate time for radiotherapy and radiotherapy sensitization,and hope that it will be helpful to the treat-ments.

Objective To clarify the relationship of p63 and CD44 expressions with age and ultraviolet irradiation in the process of skin aging.Methods Full-thickness skin samples were obtained from the normal skin at different sites,including 44 sun-exposed sites(face,neck,dorsal aspect of the forearm),46 sun-protected sites(abdomen,thigh),and 31 semi-exposed sites(back,flexor aspect of the forearm,upper arm,leg),of 121 volunteers(63 males and 58 females).Of all the volunteers,55 were 10 t0 30 vears of age (young),41 were 31 to 50 years of age(adult),and 25 were 51 to 79 years of age(old).Immunohistochemisty using antibodies to p63 and CD44 was applied to detect the expression of p63 and CD44 in these specimens.Imaging analysis software was utilized to calculate the percentage of p63-expressing cells in basal layer as well as the expression level of p63 and CD44.Results In stratum basale,the percentage of p63-expressing cells decreased with age(r=-0.218,P＜0.05),but showed no significant difference among sun-exposed skin,semi-exposed skin and sun-protected skin(P＞0.05).The correlation between the expression intensity of epidermal p63 and age was negative in sun-exposed sites(r=-0.389,P=0.010),positive in sun-protected sites(r=0.341,P＜0.05).but non-significant in semi-exposed sites(P＞0.05).With regard to the expression of epidermal p63,no significant difference was observed among sun-exposed sites,sun-protected sites and semi-exposed sites in volunteers aged from 10 to 30 years.while in those aged from 31 to 50 years and from 51 to 79 years.sun-protected sites was significantly higher than semi-exposed sites followed by sun-exposed sites(all P＜0.05).There was a weak correlation between the expression intensity of epidermal CD44 and age(r=-0.083,P＜0.05),but no significant difierence was noted among the three kinds of sites(P＞0.05).Conclusions In stratum basale,the percentage of p63.expressing cells and the expression intensity Of CD44 negatively correlate with age,but have no correlation with ultraviolet irradiation.In individuals aged from 31 to 50 years and from 51 t0 79 vears.the expression of epidermal p63 decreases with the accumulation of sun-exposure.

Objective:To analyze bone strength index, osteoporosis and fracture in ankylosing spondylitis (AS) patients with radiologic hip involvement and explore the characteristics of bone strength in these patients.Methods:According to bath ankylosing spondylitis radiology hip index (BASRI-hip) score, 339 AS patients were divided into two groups. The differences of bone strength in each group were analyzed by t-test, Mann-Whitney U test and χ2 test. Logistic regression was used to analyze the risk factors of bone strength index. The correlation between quantitative ultrasound and dualenergy X-ray absorptiometry (DXA) (total hip, g/cm 2) was analyzed. Pearson correlation analysis was used. Results:①Bone strength index of quantitative ultrasound was positively correlated with bone mineral density of DXA (total hip, g/cm 2), r=0.553, P<0.01. ② The age of 27(23, 37) years old in radiographic hip involvement was lower than 37(28, 48) years old in non-radiographic hip involvement, and the difference was statistically significant ( Z=-5.986, P<0.01). There were no differences in gender and course of disease between the two groups ( P>0.05).③ The radiographic hip involvement in AS patients whose ages were younger than 50, when compared with non-radiographic hip involvement patients, the bone strength index was lower (78±18 vs 84±16), while bone strength was lower than patients at the same age (41.0% vs 18.4%), however, the incidences of osteoporosis (42.7% vs 28.8%) and fragility fracture (3.7% vs 0%) were significantly higher ( t=3.028, P<0.01; χ2=16.758, P<0.01; χ2=5.886, P=0.019; χ2=4.67, P=0.038). For AS patients whose ages were ≥50, there were no significant differences between the two groups ( P>0.05). ④ Multivariate analysis showed that radiographic hip involvement [ OR (95% CI)=1.912(1.05, 3.48)], age [ OR (95% CI)=0.94(0.911, 0.97)] and body mass index (BMI) (kg/m 2) [ OR (95% CI) =0.875(0.807, 0.948)] were associated with lower bone strength. Conclusion:There is positive correlation between the bone strength index of quantitative ultrasound and bone mineral density of DXA. AS patients with radiographic hip involvement are characterized by decreased bone strength and are more likely to develop osteoporosis and fragile fractures. The risk factors of low bone strength are radiographic hip involvement, age and BMI.

Carbon-limited continuous culture was used to study the relationship between the growth of Aspergillus niger and the production of glucoamylase. The result showed that when the specific growth rate was lower than 0.068 h(-1), the production of glucoamylase was growth-associated, when the specific growth rate was higher than 0.068 h(-1), the production of glucoamylase was not growth-associated. Based on the result of continuous culture, the Monod dynamics model of glucose consumption of A. niger was constructed, Combining Herbert-Pirt equation of glucose and oxygen consumption with Luedeking-Piret equation of enzyme production, the black-box model of Aspergillus niger for enzyme production was established. The exponential fed-batch culture was designed to control the specific growth rate at 0.05 h(-1) by using this model and the highest yield for glucoamylase production by A. niger reached 0.127 g glucoamylase/g glucose. The black-box model constructed in this study successfully described the glucoamylase production by A. niger and the result of the model fitted the measured value well. The black-box model could guide the design and optimization of glucoamylase production by A. niger.
Aspergillus niger ; metabolism ; Batch Cell Culture Techniques ; Carbon ; Culture Media ; Glucan 1,4-alpha-Glucosidase ; biosynthesis ; Glucose ; Industrial Microbiology ; methods ; Oxygen

13 C isotopic abundance of intracellular free amino acid with a characteristic of fast- turnover can quickly reflect changes in intracellular metabolic state. But the concentration of intracellular free amino acid is low, the existed 13 C isotope detection method based on GC-MS can not satisfy the requirement with full scan mode. In this study, the selected ion monitoring method was used to detect accuracy higher likelihood of analysis of 13 C isotopic abundance of free intracellular amino acid. First, in the full scan mode we analyzed of the fracture law of different amino acids, found the feature corresponding to each amino acid fragments, and established 16 kinds of free intracellular amino acids characteristic fragment library. Then using this characteristic fragment library, only specific m/z signal was detected in sample analysis, which realized the selected ion monitoring and improved the quality of signal. The results of amino acid standards showed that the signal-to-noise ratio, measurement precision and accuracy were improved by 17, 2. 0 and 3. 8 times compared with the full scan mode. In the analysis of coenzyme Q10 producing strains of samples, this method was successfully used to detect isotopic abundance of 8 kinds of free intracellular amino acids. This method plays an important role in the detection of 13 C isotopic abundance of the intracellular free amino acid in cell metabolism research.

A method for measuring 13C isotopic abundance of intracellular metabolites of Saccharopolysporaerythraea by ultra-high performance liquid chromatography (UPLC)-triple quadrupole mass spectrometry was established.First, the chromatographic conditions of UPLC were optimized, and then the MS conditions such as unique tube lens voltage, collision energy, and ion pair were optimized.On the bases of length of the parent and daughter ions carbon chains and whether the daughter ions contain 13C atoms, the one-to-one method, one-to-many method and SIM method were established for measuring 13C isotopic abundance.Then these methods were used to measure naturally labeled intracellular metabolite standards and 13C labeled samples, and according to the gap between the experimental value and the theoretical value, the best method was established for each metabolite of different characteristics.The results showed that one-to-one method was most effective for measuring the metabolites of daughter ions not containing 13C atoms represented by sugar phosphates, one-to-many method was the best for measuring the metabolites of both parent and daughter ions containing 13C short carbon chains represented by carboxylic acids, SIM method could play a role in measuring the metabolites of both parent and daughter ions containing 13C long carbon chains represented by coenzyme A.This method had a good measurement precision and could be applied to the measurement of Saccharopolysporaerythraea intracellular metabolites, which contributed to the consequent study of metabolic mechanism and the efficient expression of erythromycin.

Objective: To understand sleep behavior among primary and middle school students and its impact on overweight and obesity changes, to provide evidence for developing obesity prevention and controlling strategies in children and adolescents. Methods: Primary and middle school students from three cities in Zhejiang Province who participated in questionnaire surveys and physical measurements in both 2017 and 2019 were selected. A follow up dataset of 605 students was developed and the relationship between sleep duration and body mass index was analyzed. Results: From 2017 to 2019, BMI Z scores for male and female participants increased by 0.24 and 0.13, respectively. BMI Z scores increased by 0.29 in students of 9-12 years old and increased by 0.11 and 0.25 in urban and rural students, respectively ( P <0.05). The prevalence of insufficient sleep duration increased from 37.0 % to 41.8% simultaneously ( χ 2=3.68, P =0.06). After adjusting for confounding factors, the BMI Z score of students with insufficient sleep was 0.20 higher than those with sufficient sleep duration ( P <0.01). Compared with participants who had sufficient sleep duration from 2017 to 2019, participants whose sleep duration changed from sufficient to insufficient, and those who always had insufficient sleep duration increased by 0.23, respectively ( P <0.05). Conclusion Insufficient sleep duration is a risk factor for obesity. Shortened sleep duration is related to weight gain, and maintaining sufficient sleep duration may reduce the risk of obesity in children and adolescents.

Aim To screen the potential inhibitors of post-transcriptional activity of pro-inflammatory media-tor TNF-α from the lipophilic constituents in Chinese Medicine Salvia miltiorrhiza Bunge ( Danshen) , we es-tablished dual luciferase reporter gene system pGL3-TNF-α3′UTR ( 3′untranslated region ) co-transfected with Renilla control gene. Methods Complementary DNA ( cDNA) template was obtained from human um-bilical vein endothelial cells ( HUVECs ) . The full length DNA of TNF-α 3′-UTR was amplified through PCR, and then connected the luciferase reporter vector pGL3-control after enzyme digestion. pGL3-TNF-α 3′UTR constructs were co-transfected with pSVRenilla into the mononuclear macrophages RAW264. 7 cells. The relative activity of reporter genes was measured by dual luciferase reporter ( DLR ) assay system after the stimulus of lipopolysaccharide ( LPS ) in presence or absence of tanshinones compounds. Results The pGL3-TNF-α3′UTR luciferase reporter gene was suc-cessfully constructed. The cloning DNA fragment and sequence were both consistent with the GENBANK da-tabase. LPS significantly induced the relative reporter activityof RAW264 . 7 cells transfected with pGL3-TNF-α 3′UTR. Among four tanshinones compounds, we found only cryptotanshinone could significantly de-crease LPS-induced relative reporter activity. Conclu-sion The pGL3-TNF-α 3′UTR construct combined with DLR assay system was successfully established, which can be used to discover the agents such as cryp-totanshinone that regulate the post-transcription of TNF-α in treatment of inflammatory and malignant dis-eases.

Objective To investigate the clinical characteristics of paraneoplastic syndrome with prominent osteoarticular involvement. Methods The clinical materials of 20 patients with paraneoplastic syndrome with prominent osteoarticular involvement were collected. The characteristics of clinical manifest-ations, laboratory tests and imagines were analyzed. Results Among the 20 patients, 16 were male and 4 were female, with a mean age of 44.5 years and a median course of 6 months. Ten cases were associated with hematological tumor and 10 cases were associated with solid tumor. Eleven cases presented as peripheral arthritis (7 cases of polyarthritis, 4 cases of oligoarthritis/monoarthritis), 5 cases presented with hypertrophic osteoarthropathy (HOA) and 4 cases presented with tumor-induced osteomalacia (TIO). Three cases were acute lymphocytic leukemia, 2 cases were multiple myeloma, 1 case was lymphoma, and 1 case was bone tumor in polyarthritis. Four oligoarthritis cases were all associated with acute lymphocytic leukemia. All 5 cases of HOA were associated with lung cancer. All 4 cases of TIO were associated with tumor of mesenchymal tissue. Extra-articular manifestations presented in 14 cases and inflammatory markers increased in 15 cases. anti-cyclic cirullinated peptide (anti-CCP) antibodies was low titer positive in only 1 case and other parameters including rheumatoid factor (RF), anti-CCP antibodies, antinuclear antibodies spectrum (ANAs) and human leukocyte antigen (HLA)-B27 were negative. Multiple bone imaging abnormalities appeared in 15 cases. Conclusion Osteoarticular manifestations may be the first symptom of malignancy and difficult to diagnose. It is necessary to be highly aware of potential malignancy.